Zhao J W, Shi G, Ping J D, Ming L
Clinical Laboratory, the First Affiliated Hospital of Zhengzhou University, Key Clinical Laboratory of Henan Province, Zhengzhou 450052, China.
Zhonghua Yi Xue Za Zhi. 2018 Sep 25;98(36):2889-2893. doi: 10.3760/cma.j.issn.0376-2491.2018.36.003.
To investigate the expression of tumor necrosis factor alpha-inducible protein 8 like-2 (TIPE2) and tissue factor (TF) in patients with bronchial asthma. And to explore the regulation of TIPE2 on TF. Sixty-five asthmatic patients and 40 healthy controls were selected from the First Affiliated Hospital of Zhengzhou University from July to November, 2017. The expression of TIPE2 and TF in peripheral blood mononuclear cells of asthmatic patients and healthy controls were detected by Western blot.The level of TF protein in plasma was detected by enzyme-linked immunosorbent assay. The changes of TIPE2 and TF mRNA expression in THP-1 cells stimulated by house dust mite extract were detected by real-time fluorescence quantitative PCR (RT-PCR). The recombinant adenovirus Adv-TIPE2 was constructed and transfected into THP-1 cells and the effect of over-expression TIPE2 on TF expression in THP-1 cells was detected by RT-PCR. The relative level of TIPE2 protein in asthmatic patients and healthy controls was 0.025±0.010 and 0.087±0.070, while that of TF was 0.40±0.27 and 0.15±0.10, respectively. Compared with healthy controls, the levels of TIPE2 protein decreased and TF protein increased in asthmatic patients, the differences were statistically significant (=-5.06, 9.04, <0.05) . TIPE2 protein level was negatively correlated with TF protein level (=-0.460 3, <0.05). The house dust mite extract reduced the expression level of TIPE2 mRNA in THP-1 cells, but increased the level of TF mRNA expression. When the concentration was 1 μg/ml, the change of TIPE2 mRNA was the most obvious at 4 h (<0.05). The recombinant adenovirus Adv-TIPE2 was successfully constructed. The level of TF mRNA expression in THP-1 cells over-expressing TIPE2 gene was reduced (<0.05). TIPE2, a negative regulator of inflammation, has a negative control effect on TF, and may be involved in the hyper-coagulable state of bronchial asthma by regulating TF expression.
探讨支气管哮喘患者肿瘤坏死因子α诱导蛋白8样2（TIPE2）和组织因子（TF）的表达情况，并探究TIPE2对TF的调控作用。选取2017年7月至11月郑州大学第一附属医院的65例哮喘患者和40例健康对照者。采用蛋白质免疫印迹法检测哮喘患者和健康对照者外周血单个核细胞中TIPE2和TF的表达。采用酶联免疫吸附测定法检测血浆中TF蛋白水平。采用实时荧光定量聚合酶链反应（RT-PCR）检测尘螨提取物刺激后THP-1细胞中TIPE2和TF mRNA表达的变化。构建重组腺病毒Adv-TIPE2并转染至THP-1细胞，采用RT-PCR检测过表达TIPE2对THP-1细胞中TF表达的影响。哮喘患者和健康对照者TIPE2蛋白相对水平分别为0.025±0.010和0.087±0.070，TF相对水平分别为0.40±0.27和0.15±0.10。与健康对照者相比，哮喘患者TIPE2蛋白水平降低，TF蛋白水平升高，差异有统计学意义（=-5.06，9.04，<0.05）。TIPE2蛋白水平与TF蛋白水平呈负相关（=-0.460 3，<0.05）。尘螨提取物降低了THP-1细胞中TIPE2 mRNA表达水平，但提高了TF mRNA表达水平。当浓度为1 μg/ml时，4 h时TIPE2 mRNA变化最明显（<0.05）。成功构建重组腺病毒Adv-TIPE2。过表达TIPE2基因的THP-1细胞中TF mRNA表达水平降低（<0.05）。TIPE2作为一种炎症负调控因子，对TF具有负向调控作用，可能通过调节TF表达参与支气管哮喘的高凝状态。
